goat polyclonal anti mouse tf antibody af3178  (R&D Systems)

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in a mouse cell line using 4 different commercial anti-mouse TF antibodies. Proteins in cell lysates (10 μg) from the mouse pancreatic cancer cell lines KPC2 wild-type (WT) and KPC2 TF knockout (KO) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a membrane. TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody. These anti-TF antibodies were detected using species-specific secondary antibodies . β-Actin was used as a loading control and detected with an anti–β-actin antibody . The binding of the anti–β-actin antibody was detected with a horse anti-mouse immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the blots were exposed for 1 second for both TF and β-actin. All blots were visualized using the iBright FL1000 imaging system. kDa, kilodalton.

Article Snippet: We evaluated the ability of the following commercially available antibodies to detect TF in KPC2 WT cells: goat polyclonal anti-mouse TF antibody AF3178 (R&D Systems), rabbit monoclonal anti-mouse TF antibody ab189483 (Abcam), rabbit monoclonal anti-mouse TF antibody 44861 (Cell Signaling Technology), and mouse monoclonal anti-mouse TF antibody H-9 (Santa Cruz; ).

Techniques: Knock-Out, Polyacrylamide Gel Electrophoresis, Membrane, Control, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the R&D Systems antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody. The binding of the R&D Systems anti-TF antibody was detected with a rabbit anti-goat immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for 1 second. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: We evaluated the ability of the following commercially available antibodies to detect TF in KPC2 WT cells: goat polyclonal anti-mouse TF antibody AF3178 (R&D Systems), rabbit monoclonal anti-mouse TF antibody ab189483 (Abcam), rabbit monoclonal anti-mouse TF antibody 44861 (Cell Signaling Technology), and mouse monoclonal anti-mouse TF antibody H-9 (Santa Cruz; ).

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in a mouse cell line using 4 different commercial anti-mouse TF antibodies. Proteins in cell lysates (10 μg) from the mouse pancreatic cancer cell lines KPC2 wild-type (WT) and KPC2 TF knockout (KO) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a membrane. TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody. These anti-TF antibodies were detected using species-specific secondary antibodies . β-Actin was used as a loading control and detected with an anti–β-actin antibody . The binding of the anti–β-actin antibody was detected with a horse anti-mouse immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the blots were exposed for 1 second for both TF and β-actin. All blots were visualized using the iBright FL1000 imaging system. kDa, kilodalton.

Article Snippet: TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody.

Techniques: Knock-Out, Polyacrylamide Gel Electrophoresis, Membrane, Control, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the R&D Systems antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody. The binding of the R&D Systems anti-TF antibody was detected with a rabbit anti-goat immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for 1 second. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody.

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in a mouse cell line using 4 different commercial anti-mouse TF antibodies. Proteins in cell lysates (10 μg) from the mouse pancreatic cancer cell lines KPC2 wild-type (WT) and KPC2 TF knockout (KO) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a membrane. TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody. These anti-TF antibodies were detected using species-specific secondary antibodies . β-Actin was used as a loading control and detected with an anti–β-actin antibody . The binding of the anti–β-actin antibody was detected with a horse anti-mouse immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the blots were exposed for 1 second for both TF and β-actin. All blots were visualized using the iBright FL1000 imaging system. kDa, kilodalton.

Article Snippet: We used the goat anti-mouse TF polyclonal antibody AF3178 (R&D Systems), the rabbit anti-mouse TF monoclonal antibody ab189483 (Abcam), the rabbit anti-mouse TF monoclonal antibody 44861 (Cell Signaling Technology), and the mouse anti-mouse TF monoclonal antibody H9 (Santa Cruz Biotechnology) to measure TF in the mouse pancreatic cancer cell line KPC2 wild-type and KPC2 TF knockout cell lysates, as well as various tissues from wild-type and low TF mice (no mouse TF expression) using western blotting.

Techniques: Knock-Out, Polyacrylamide Gel Electrophoresis, Membrane, Control, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the R&D Systems antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody. The binding of the R&D Systems anti-TF antibody was detected with a rabbit anti-goat immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for 1 second. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: We used the goat anti-mouse TF polyclonal antibody AF3178 (R&D Systems), the rabbit anti-mouse TF monoclonal antibody ab189483 (Abcam), the rabbit anti-mouse TF monoclonal antibody 44861 (Cell Signaling Technology), and the mouse anti-mouse TF monoclonal antibody H9 (Santa Cruz Biotechnology) to measure TF in the mouse pancreatic cancer cell line KPC2 wild-type and KPC2 TF knockout cell lysates, as well as various tissues from wild-type and low TF mice (no mouse TF expression) using western blotting.

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Optimization of a Western blot protocol for the detection of low levels of tissue factor in human cells

doi: 10.1016/j.rpth.2025.103016

Figure Lengend Snippet: Effect of different secondary antibodies on the detection of tissue factor (TF) by Western blotting using the same primary antibody. Cell lysates from HAP-1 wild-type (WT) and HAP-1 TF knockout (KO; both 40 μg) and recombinant TF (rTF; Innovin, 500 pg, Thermo Fisher Scientific, catalog [cat] number 10873566) were run on a gel, and proteins were transferred to a membrane. The blots were blocked with 5% nonfat milk (Walmart, nonfat dry milk powder) in Tris-buffered saline (TBS)-Tween. TF was detected using a goat anti-TF polyclonal antibody (R&D Systems, cat number AF2339) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. Anti-TF antibody binding was detected using either a mouse antigoat horseradish peroxidase (HRP)-linked secondary antibody (Santa Cruz, cat number sc-2354), a rabbit antigoat HRP-linked secondary antibody (Thermo Fisher Scientific, cat number 31402), or a donkey antigoat HRP-linked secondary antibody (Thermo Fisher Scientific, cat number A15999), all diluted 1:5000 in blocking buffer. The membranes were incubated with Clarity Max Western Enhanced Chemiluminescence Substrate (Bio-Rad, cat number 1705062) for 10 seconds. β-Actin was used as a loading control and was detected using a mouse anti–β-actin antibody (Santa Cruz, cat number sc-47778) diluted 1:5000 in 5% bovine serum albumin in TBS-Tween. An HRP-linked horse anti-mouse HRP antibody (Cell Signaling Technologies, cat number 7076) diluted 1:5000 in blocking buffer was used to detect anti–β-actin binding. Clarity Western Enhanced Chemiluminescence Substrate was added for 10 seconds prior to imaging. For imaging, the blot was exposed for 5 seconds (TF) and 10 seconds (β-actin). The blots were visualized using the iBright FL1000 imaging system (Thermo Fisher Scientific). kDa, kilodalton.

Article Snippet: TF was detected using a goat anti-TF polyclonal antibody (R&D Systems, cat number AF2339) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween.

Techniques: Western Blot, Knock-Out, Recombinant, Membrane, Saline, Binding Assay, Blocking Assay, Incubation, Control, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Optimization of a Western blot protocol for the detection of low levels of tissue factor in human cells

doi: 10.1016/j.rpth.2025.103016

Figure Lengend Snippet: Detection of tissue factor (TF) in different cell lines using 4 different anti-TF antibodies. Cell lysates (BxPC-3 [1 μg] and THP-1 control; THP-1 + lipopolysaccharide [LPS]; HAP-1 wild-type [WT]; and HAP-1 TF knockout [KO; all 40 μg]) were run on separate gels and transferred to membranes. TF was detected using 1 of 4 antibodies: (A) Novus NBP2-15139 (rabbit polyclonal), (B) R&D AF2339 (goat polyclonal), and (C) Abcam ab252918 (lot number 1096425-7, designated Abcam 1, rabbit monoclonal) and (D) Abcam ab228968 (designated Abcam 2) diluted 1:1000 in 5% bovine serum albumin in Tris-buffered saline (TBS)-Tween. The blots were blocked with 5% nonfat milk (Walmart, nonfat dry milk powder) in TBS-Tween. The binding of the rabbit anti-TF antibodies was detected using an HRP-linked goat anti-rabbit secondary antibody (Thermo Fisher Scientific, cat number 31460), and the binding of the goat anti-TF antibody was detected using an HRP-linked rabbit antigoat secondary antibody (Thermo Fisher Scientific, cat number 31402). Both secondary antibodies were diluted 1:5000 in 5% milk in TBS-Tween. The membranes were incubated with Clarity Max Western Enhanced Chemiluminescence Substrate (Bio-Rad, cat number 1705062) for 10 seconds. α-Actinin was used as a loading control and detected using a rabbit anti–α-actinin antibody (Cell Signaling Technologies, cat number 3134) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. The binding of the rabbit anti–α-actinin antibody was detected using an HRP-linked goat anti-rabbit secondary (Thermo Fisher Scientific, cat number 31460) diluted 1:5000 in blocking buffer. Clarity Western Enhanced Chemiluminescence Substrate was added for 10 seconds prior to imaging. For imaging, the blots were exposed for 10 seconds (A and B) or 2 seconds for TF detection (C and D) . All blots were exposed for 10 seconds for α-actinin detection. The blots were visualized using the iBright FL1000 imaging system (Thermo Fisher Scientific). For α-actinin, the brightness of the entire blot was adjusted. kDa, kilodalton.

Article Snippet: TF was detected using a goat anti-TF polyclonal antibody (R&D Systems, cat number AF2339) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween.

Techniques: Control, Knock-Out, Saline, Binding Assay, Incubation, Western Blot, Blocking Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Optimization of a Western blot protocol for the detection of low levels of tissue factor in human cells

doi: 10.1016/j.rpth.2025.103016

Figure Lengend Snippet: Effect of different secondary antibodies on the detection of tissue factor (TF) by Western blotting using the same primary antibody. Cell lysates from HAP-1 wild-type (WT) and HAP-1 TF knockout (KO; both 40 μg) and recombinant TF (rTF; Innovin, 500 pg, Thermo Fisher Scientific, catalog [cat] number 10873566) were run on a gel, and proteins were transferred to a membrane. The blots were blocked with 5% nonfat milk (Walmart, nonfat dry milk powder) in Tris-buffered saline (TBS)-Tween. TF was detected using a goat anti-TF polyclonal antibody (R&D Systems, cat number AF2339) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. Anti-TF antibody binding was detected using either a mouse antigoat horseradish peroxidase (HRP)-linked secondary antibody (Santa Cruz, cat number sc-2354), a rabbit antigoat HRP-linked secondary antibody (Thermo Fisher Scientific, cat number 31402), or a donkey antigoat HRP-linked secondary antibody (Thermo Fisher Scientific, cat number A15999), all diluted 1:5000 in blocking buffer. The membranes were incubated with Clarity Max Western Enhanced Chemiluminescence Substrate (Bio-Rad, cat number 1705062) for 10 seconds. β-Actin was used as a loading control and was detected using a mouse anti–β-actin antibody (Santa Cruz, cat number sc-47778) diluted 1:5000 in 5% bovine serum albumin in TBS-Tween. An HRP-linked horse anti-mouse HRP antibody (Cell Signaling Technologies, cat number 7076) diluted 1:5000 in blocking buffer was used to detect anti–β-actin binding. Clarity Western Enhanced Chemiluminescence Substrate was added for 10 seconds prior to imaging. For imaging, the blot was exposed for 5 seconds (TF) and 10 seconds (β-actin). The blots were visualized using the iBright FL1000 imaging system (Thermo Fisher Scientific). kDa, kilodalton.

Article Snippet: We examined the ability of 3 different antibodies to detect TF in low-expressing cell lines: rabbit polyclonal anti-human TF antibody NBP2-15139 (Novus Biologicals), goat polyclonal anti-human TF antibody AF2339 (R&D Systems), and rabbit monoclonal anti-human TF antibody ab252918 (clone EPR22548-240; Abcam).

Techniques: Western Blot, Knock-Out, Recombinant, Membrane, Saline, Binding Assay, Blocking Assay, Incubation, Control, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Optimization of a Western blot protocol for the detection of low levels of tissue factor in human cells

doi: 10.1016/j.rpth.2025.103016

Figure Lengend Snippet: Detection of tissue factor (TF) in different cell lines using 4 different anti-TF antibodies. Cell lysates (BxPC-3 [1 μg] and THP-1 control; THP-1 + lipopolysaccharide [LPS]; HAP-1 wild-type [WT]; and HAP-1 TF knockout [KO; all 40 μg]) were run on separate gels and transferred to membranes. TF was detected using 1 of 4 antibodies: (A) Novus NBP2-15139 (rabbit polyclonal), (B) R&D AF2339 (goat polyclonal), and (C) Abcam ab252918 (lot number 1096425-7, designated Abcam 1, rabbit monoclonal) and (D) Abcam ab228968 (designated Abcam 2) diluted 1:1000 in 5% bovine serum albumin in Tris-buffered saline (TBS)-Tween. The blots were blocked with 5% nonfat milk (Walmart, nonfat dry milk powder) in TBS-Tween. The binding of the rabbit anti-TF antibodies was detected using an HRP-linked goat anti-rabbit secondary antibody (Thermo Fisher Scientific, cat number 31460), and the binding of the goat anti-TF antibody was detected using an HRP-linked rabbit antigoat secondary antibody (Thermo Fisher Scientific, cat number 31402). Both secondary antibodies were diluted 1:5000 in 5% milk in TBS-Tween. The membranes were incubated with Clarity Max Western Enhanced Chemiluminescence Substrate (Bio-Rad, cat number 1705062) for 10 seconds. α-Actinin was used as a loading control and detected using a rabbit anti–α-actinin antibody (Cell Signaling Technologies, cat number 3134) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. The binding of the rabbit anti–α-actinin antibody was detected using an HRP-linked goat anti-rabbit secondary (Thermo Fisher Scientific, cat number 31460) diluted 1:5000 in blocking buffer. Clarity Western Enhanced Chemiluminescence Substrate was added for 10 seconds prior to imaging. For imaging, the blots were exposed for 10 seconds (A and B) or 2 seconds for TF detection (C and D) . All blots were exposed for 10 seconds for α-actinin detection. The blots were visualized using the iBright FL1000 imaging system (Thermo Fisher Scientific). For α-actinin, the brightness of the entire blot was adjusted. kDa, kilodalton.

Article Snippet: We examined the ability of 3 different antibodies to detect TF in low-expressing cell lines: rabbit polyclonal anti-human TF antibody NBP2-15139 (Novus Biologicals), goat polyclonal anti-human TF antibody AF2339 (R&D Systems), and rabbit monoclonal anti-human TF antibody ab252918 (clone EPR22548-240; Abcam).

Techniques: Control, Knock-Out, Saline, Binding Assay, Incubation, Western Blot, Blocking Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Optimization of a Western blot protocol for the detection of low levels of tissue factor in human cells

doi: 10.1016/j.rpth.2025.103016

Figure Lengend Snippet: Effect of deglycosylation on lysates from BxPC-3, HAP-1 wild-type (WT), HAP-1 tissue factor (TF) knockout (KO) cells, and lipopolysaccharide (LPS)-stimulated THP-1 cells. Cell lysates (BxPC-3 [1 μg]; HAP-1 WT and HAP-1 TF KO; and LPS-stimulated THP-1 cells [all 40 μg]) were treated with no enzyme or PNGase F. The lysates from the LPS-stimulated THP-1 cells were run on a separate gel from the other lysates. After running the gels, the proteins were transferred to membranes. The blots were blocked with 5% nonfat milk (Walmart, nonfat dry milk powder) in Tris-buffered saline (TBS)-Tween. TF (glycosylated TF [TF] and deglycosylated TF [dg-TF]) was detected using 1 of 3 antibodies: (A) Novus NBP2-15139, (B) R&D AF2339, and (C) Abcam ab252918 (lot number GR3270793-2, designated Abcam 1) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. The binding of the rabbit anti-TF antibodies was detected using an HRP-linked goat anti-rabbit secondary antibody (Thermo Fisher Scientific, cat number 31460), and the binding of the goat anti-TF antibody was detected using an HRP-linked rabbit antigoat secondary antibody (Thermo Fisher Scientific, cat number 31402). Both secondary antibodies were diluted 1:5000 in 5% milk in TBS-Tween. The membranes were incubated with Clarity Max Western Enhanced Chemiluminescence Substrate (Bio-Rad, cat number 1705062) for 10 seconds. α-Actinin was used as a loading control and detected using a rabbit anti–α-actinin antibody (Cell Signaling Technologies, cat number 3134) diluted 1:1000 in 5% bovine serum albumin in TBS-Tween. The binding of the rabbit anti–α-actinin antibody was detected using an HRP-linked goat antir-abbit secondary antibody (Thermo Fisher Scientific, cat number 31460) diluted 1:5000 in blocking buffer. The membranes were incubated with Clarity Western Enhanced Chemiluminescence Substrate for 10 seconds. For imaging, the blots were exposed for 2 seconds for TF detection and 10 seconds for α-actinin detection. The blots were visualized using the iBright FL1000 imaging system (Thermo Fisher Scientific). For α-actinin, the brightness of the entire blot was adjusted. kDa, kilodalton; NS, non-specifc.

Article Snippet: We examined the ability of 3 different antibodies to detect TF in low-expressing cell lines: rabbit polyclonal anti-human TF antibody NBP2-15139 (Novus Biologicals), goat polyclonal anti-human TF antibody AF2339 (R&D Systems), and rabbit monoclonal anti-human TF antibody ab252918 (clone EPR22548-240; Abcam).

Techniques: Knock-Out, Saline, Binding Assay, Incubation, Western Blot, Control, Blocking Assay, Imaging